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为探索COⅠ基因条形码技术在龟鳖目物种分类与鉴定中的适用性,本研究应用通用引物PCR扩增获得龟鳖目动物9科26属45种共205个样品的634 bp COⅠ基因序列片段进行研究分析。结果显示,COⅠ基因在龟鳖目动物中存在少量的碱基插入和缺失,共计12种20个位点,物种占比34.28%;平均碱基含量(A+T)为55.42%,显著高于(G+C)的44.58%。通过Kimura's 2-parameter计算45个物种的种内遗传距离为0～0.023 98,平均为0.003 2;种间遗传距离为0.022 3～0.334 9,平均为0.1 637,种间平均遗传距离约为种内平均遗传距离的51.16倍;有3个物种的种内遗传距离大于Hebert推荐区分物种的最小种间遗传距离0. 02,其中1个物种的种内遗传距离大于45个物种的种间最小遗传距离0.022 3。同科内不同属间及同目内不同科间的遗传距离分别为0.146 7、0.212 7,显示物种间遗传距离随着分类阶元的上升而逐步扩大。系统进化树聚类结果显示,不同个体依据亲缘关系程度不同分别聚为独立的一支,没有出现物种交叉分布,能准确反映个体间的亲缘关系。结果表明利用COⅠ基因鉴定龟鳖目物种具有可行性,但科内属间及以上分类阶元进化节点自展率不高,无法通过COⅠ基因明确其进化关系,须结合其他DNA指标加以确认。
Abstract：In order to explore the applicability of COⅠgene barcoding technique in the classification and identification of turtle species. Universal primers PCR amplification was used to analyze 634 bp CO Ⅰ gene sequence fragments, which was obtained from 205 samples of 45 species belong to 9 families and 26 genuses of Trionyx.The results showed that the COⅠgenes had a small number of base insertions and deletions in turtles, total 20 sites in 12 species, accounted for 34.28% of all species; The average base content of A +T was 55.42%, significantly higher than 44.58% of G+C. Calculated with Kimura's 2-parameter model, the intraspecific genetic distances of 45 species ranged from 0 to 0.0239 8, with an average of 0.003 2, and interspecific genetic distance ranged from0.022 3 to 0.334 9, with an average of 0.163 7. The interspecific genetic distances were about 51.16 times higher than the intraspecies genetic distances. There were three species in which the intraspecific genetic distances were greater than Hebert recommended value to distinguish species of the minimum genetic distance of 0.02, one intraspecific genetic distance was greater than the minimum interspecific genetic distance of 0.022 3 in 45 species.The genetic distance between different genuses within family and different families within order was 0.146 7 and 0.212 7,respectively, indicating that the genetic distances between species increases gradually with the increase of taxonomic category. The results of phylogenetic tree clustering showed that different individuals are grouped into independent branches according to the degree of genetic relationship. There is no cross distribution which can accurately reflect the relationship between species. The results also showed that using COⅠgene to identify the species is feasible.However, the self-expansion rate of evolution node between genuses within family and above is not high. The evolutionary relationship can not be determined by COⅠgene, which must be confirmed by other DNA indexes.
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基本信息:
DOI：10.13417/j.gab.038.003935
中图分类号:S917.4
引用信息:
[1]叶朝阳,李伟,赵建,等.基于COⅠ基因的龟鳖目动物鉴定与分类进化分析[J].基因组学与应用生物学,2019,38(09):3935-3945.DOI:10.13417/j.gab.038.003935.
基金信息:
中央级公益性科研院所基本科研业务费专项资金资助(2014A11);; 科技基础性工作专项资助(2013FY110700)共同资助
2018-08-02
2018-08-02
2018-08-02